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PubMed1986

Spatial and temporal pattern of hsp26 expression during normal development.

The tissue-specific patterns of developmental expression of hsp26-lacZ fusion genes inserted into Drosophila melanogaster by germline transformation were analyzed in several transformant lines utilizing a histochemical assay for beta-galactosidase activity on whole animals. We compared this pattern to the tissue-specific distribution of endogenous hsp26 RNA determined using hybridization of probes to RNA in situ in tissue sections. Both assays reveal that hsp26 is expressed in numerous tissues during development including spermatocytes, nurse cells, epithelium, imaginal discs, proventriculus and neurocytes. The ease and resolution of the whole-animal beta-galactosidase assay makes it particularly attractive for the elucidation of sequences involved in such complex regulation. The original hsp26-lacZ fusion gene contained 2 kb of sequence upstream of the transcription start. A construct containing only 278 bp upstream was still expressed in spermatocytes but no longer in nurse cells. In a few instances, the fusion genes were expressed in tissues for which there was no evidence for expression of the endogenous hsp26 gene. These novel patterns appear to be a result of chromosomal position since they were observed in only one or a subset of transformant lines containing identical inserts.

Animals↗
PubMed1984

Gastrointestinal hormones in birds: morphological, chemical, and developmental aspects.

Historically, the enterochromaffin cell was the first endocrine cell type detected in avian gut; subsequently, a number of types of such cells were distinguished on the basis of the ultrastructural features of the secretory granules. More recently, immunocytochemical procedures have revealed somatostatin-, pancreatic polypeptide (PP)-, polypeptide YY-, glucagon-, secretin-, vasoactive intestinal peptide (VIP)-, gastrin-, cholecystokinin-, neurotensin-, bombesin-, substance P-, enkephalin-, motilin-, and FMRFamide-like immunoreactivity in avian gastrointestinal endocrine cells. Most endocrine cells are located in the antrum; there are a number in the proventriculus and small intestine but few in the gizzard, cecum, and rectum. Several avian gastroenteropancreatic hormones, including glucagon, VIP, secretin, bombesin, neurotensin, and PP, have been isolated and sequenced. They resemble the equivalent mammalian peptides in terms of molecular size but differ in amino acid composition and sequence; some (e.g., VIP) differ only in minor respects, others (e.g., secretin) more radically. Gastrointestinal endocrine cells appear late in development; available data indicate that few types are recognized by either immunocytochemistry or electron microscopy before 16 days of incubation. Experimental evidence has shown that at least the majority of gut endocrine cells are of endodermal origin and are not derived from the neural crest or neuroectoderm as earlier proposed. In early embryos, the progenitors of gastrointestinal endocrine cells are more widespread than are the differentiated cells in chicks at hatching. This, along with other observations, raises the question of factors that might influence the differentiation of gut endocrine cells.

APUD Cells↗
PubMed1994

The development of cellular junctions in the Drosophila embryo.

The pattern and development of cellular junctions in the different tissues of the Drosophila embryo from the blastoderm stage until hatching were analyzed. The cellular junctions found include: gap junctions, two types of septate junctions, and several types of cell-cell and cell-substrate adherens junctions. During early and mid embryogenesis (stages 4 to 13) only spot adherens junctions, gap junctions, and zonulae adherentes prevail. Scattered spot adherens junctions are already formed at the blastoderm stage. During and shortly after gastrulation, spot adherens junctions become concentrated at the apical pole and fuse into continuous zonulae adherentes in the posterior endoderm and the ectoderm. In addition to the zonulae adherentes, ectodermally derived epithelia possess scattered gap junctions and form pleated septate junctions and hemiadherens junctions during late embryogenesis (stages 14 to 17). Mesenchymal tissues (i.e., all nonepithelial tissues of the embryo, including the neural primordium and, transiently, the mesoderm and endoderm) possess both spot adherens junctions and gap junctions at a low frequency. Initially, the midgut epithelium does not establish a junctional complex and possess only gap junctions and spot adherens junctions. Only late in development does a circumferential smooth septate junction develop; zonulae adherentes are missing. The various derivatives of the mesoderm express spot adherens junctions, hemiadherens junctions, and gap junctions, but never zonulae adherentes or septate junctions. After organogenesis, several different types of tissue-specific adherens junctions are formed, among them connecting hemiadherens junctions (between gut epithelium and visceral muscle and early during the formation of the muscle tendon junction), muscle tendon junctions (between somatic muscle and tendon cells), fasciae adherentes (between the cells of both the visceral muscle and the dorsal vessel), and autocellular nephrocyte junctions (in nephrocytes). Interesting exceptions to the general pattern of junctional development are provided by the outer epithelial layer of the proventriculus and the Malpighian tubules. Both tissues develop as typical ectodermal epithelia and possess zonulae adherentes. During late embryogenesis, both epithelia lose the zonulae adherentes and form smooth rather than pleated septate junctions, thereby expressing a junctional complex similar to that of the endodermally derived midgut epithelium.

Animals↗
PubMed2002

Steroid regulation of midgut cell death during Drosophila development.

Steroid hormones trigger dynamic tissue changes during animal development by activating cell proliferation, cell differentiation, and cell death. Here we characterize steroid regulation of changes in midgut structure during the onset of Drosophila metamorphosis. Following an increase in the steroid 20-hydroxyecdysone (ecdysone) at the end of larval development, future adult midgut epithelium is formed, and the larval midgut is rapidly destroyed. Mutations in the steroid-regulated genes BR-C and E93 differentially impact larval midgut cell death but do not affect the formation of adult midgut epithelia. In contrast, mutations in the ecdysone-regulated E74A and E74B genes do not appear to perturb midgut development during metamorphosis. Larval midgut cells possess vacuoles that contain cellular organelles, indicating that these cells die by autophagy. While mutations in the BR-C, E74, and E93 genes do not impact DNA degradation during this cell death, mutations in BR-C inhibit destruction of larval midgut structures, including the proventriculus and gastric caeca, and E93 mutants exhibit decreased formation of autophagic vacuoles. Dying midguts express the rpr, hid, ark, dronc, and crq cell death genes, suggesting that the core cell death machinery is involved in larval midgut cell death. The transcription of rpr, hid, and crq are altered in BR-C mutants, and E93 mutants possess altered transcription of the caspase dronc, providing a mechanism for the disruption of midgut cell death in these mutant animals. These studies indicate that ecdysone triggers a two-step hierarchy composed of steroid-induced regulatory genes and apoptosis genes that, in turn, regulate the autophagic death of midgut cells during development.

Animals↗
PubMed1995

Distribution of NADPH-diaphorase activity in the embryonic chicken gut.

The appearance and distribution of NADPH-diaphorase activity in neuronal cells and fibres in different regions of the embryonic chicken gut was studied histochemically using whole mount preparations and cryostat sections. NADPH-diaphorase activity was detected in neuronal cell bodies as early as embryonic day 5.5 (E5.5 - the earliest age examined), mainly in the foregut, although some positive cells were also seen in the hindgut at this stage. NADPH-diaphorase-positive fibres were first detected in the developing nerve tracts which connect the ganglia at E5.5. The complexity of the network was maximal in the proventriculus-gizzard junction. By E9.5, NADPH-diaphorase-positive fibres were found in the circular muscle layer. NADPH-diaphorase-positive submucosal neurons were first detected at E11.5. The density of innervation was maximal at E15.5 and declined later development. The expression of neuronal NADPH-diaphorase activity progressed in a craniocaudal direction and followed a developmental pattern similar to that previously described for several neuropeptides in the avian gut.

Animals↗
PubMed1981

An immunocytochemical survey of endocrine cells in the gastrointestinal tract of chicks at hatching.

The distribution of gastrin-, cholecystokinin-, glucagon-, secretin-, vasoactive intestinal polypeptide-, substance P-, bombesin-, neurotensin-, motilin-, somatostatin- and avian pancreatic polypeptide-like cells, demonstrated by indirect immunocytochemistry, was studied in samples from the following regions: proventriculus, gizzard, pylorus, duodenum, upper and lower ileum, caeca and rectum. The pylorus is particularly rich in gastrin-, neurotensin- and somatostatin-like cells. No cells immunoreactive for gastric inhibitory polypeptide or insulin were detected. In a number of instances the same cells were found to stain with antisera raised to different gut peptides. This happened with antisera detecting gastrin- and neurotensin-like cells, with secretin, vasoactive intestinal polypeptide, glucagon and substance P. The possibility that antigenic determinants to more than one peptide are contained in certain endocrine-like cells is considered.

Animals↗
PubMed1976

Endocrine cells of the stomach of chicks around the time of hatching.

The proventriculus, gizzard and pyloric antrum (region between the gizzard and the duodenum) of 18-day Black Australorp chick embryos and of chicks within 30 h of hatching have been studied by electron microscopy. D and EC cells, and putative G, D1 and A-like cells were identified (terminology of Solcia et al., 1973) but no ECL cells. No endocrine cells of any kind were revealed in the gizzard.

Animals↗
PubMed1983

Immunocytochemical identification of alpha-endorphin-like material in neurones of the brain and corpus cardiacum of the blowfly, Calliphora vomitoria (Diptera).

A group of the 24-26 paraldehyde fuchsin-positive median neurosecretory cells (MNC) in the pars intercerebralis of the brain of the blowfly, Calliphora vomitoria, has shown immunoreactivity towards three different antibodies to alpha-endorphin, a peptide that corresponds to the amino acid sequence present between residues 61 and 76 of the precursor molecule, beta-lipotropin (beta-LPH). The immunoreactive material could be followed in axons within the median bundle, the tract through which neurosecretory material from the MNC is passed down to the corpus cardiacum (CC). The alpha-endorphin-immunoreactive material was observed leaving the CC in the cardiac-recurrent nerve, dorsal to the proventriculus, in the direction of the abdomen. The cells that contain the alpha-endorphin-like material are different from those of the MNC that contain insulin-, pancreatic polypeptide-, and gastrin/CCK-like peptides. This finding demonstrates the considerable complexity and peptidergic nature of the MNC and constitutes further evidence that morphinomimetic-like peptides are present in the nervous system of invertebrates.

Animals↗
PubMed1992

Tissue-specific position effects on alcohol dehydrogenase expression in Drosophila melanogaster.

Twenty transformed lines have been isolated as a result of the germ line insertion of a 3.2 kb alcohol dehydrogenase (Adh) gene fragment into an Adh negative strain of Drosophila melanogaster by P element-mediated transformation. More than half of these lines exhibited abnormal ADH expression. The level of ADH expression ranges from zero in some lines to near normal levels in others, and the pattern of ADH expression in the larval gut is also abnormal in many of these lines. Each of the abnormal tissue-specific patterns is stable and characterized by the absence or reduction of ADH expression in certain tissues. High levels of ectopic expression were not observed. In two of these lines, the pattern of ADH staining is highly restricted: it is limited to the medial midgut in line MM-50, and to the gastric caecae and the proventriculus in line GC-1. In heterozygotes between these two lines ADH is expressed in both of these tissues. To test the hypothesis that this abnormal expression is due to position effects, inserts were mobilized to new locations. The mobilized inserts exhibited new patterns of tissue-specific expression associated with new cytological insert locations, showing that the abnormal expression in lines MM-50 and GC-1 is due to tissue-specific position effects and not to mutations. The results are discussed in the context of chromatin structure as a possible cause of these position effects.

Alcohol Dehydrogenase↗
PubMed1982

Localization of bombesin and GRP (gastrin releasing peptide) sequences in gut nerves or endocrine cells.

Five anti-gastrin releasing peptide (GRP) sera have been characterized against GRP, bombesin and related polypeptides spotted on cellulose acetate discs. Antibodies reacting with the C-terminal G-14 sequence of bombesin and the 19-27 sequence of GRP, were detected in all sera. Antibodies directed exclusively against the bombesin unrelated 1-17 sequence of GRP were found only in one serum (R-6902). With parallel immunohistochemical tests only the C-terminal immunoreactivity was detected in endocrine-paracrine cells of the chicken proventriculus, while both immunoreactivities were present in nerve fibres and a few nerve cell bodies of the mammalian gut. The distribution of GRP- and bombesin-like immunoreactive nerves in the gastric mucosa of both pyloric and oxyntic type the submucosal and myenteric plexus along the whole gastrointestinal wall and at sphincter regions is detailed.

Animals↗
PubMed1984

Immunocytochemical localization of gastrin-releasing peptide/bombesin-like immunoreactive neurons in insects.

GRP/bombesin-like immunoreactive material was immunocytochemically detected in neurons of seven insect species belonging to seven orders, while such neurons were not found in three insect species belonging to two other orders. In some insect species certain neurons were found in corresponding places and approximately the same numbers. It seems likely that such neurons have a common evolutionary origin and are homologous. The fact that the GRP-antiserum reveals such homologous neurons in species belonging to different orders, suggests that the part of the GRP/bombesin-like peptide recognized by the antiserum has been relatively stable during evolution. As the GRP-antiserum had to be used in much higher concentrations on insect tissue than for GRP endocrine cells in chicken proventriculus, the chemical resemblance of the insect peptide(s) to GRP and bombesin may be limited.

Animals↗
PubMed1986

Histamine in endocrine cells in the stomach. A survey of several species using a panel of histamine antibodies.

Antibodies to histamine were used to examine the localization of the amine in cells of the stomach and upper small intestine of a great variety of species, including cartilaginous and bony fish, amphibia, reptiles (lizard), birds (chicken) and a large number of mammals. In all species gastric histamine was localized in endocrine cells (invariably found in the epithelium) and mast cells (usually with an extra-epithelial localization). The endocrine cells were identified as such by immunostaining with antibodies to chromogranin A and the mast cells were identified by toluidine blue staining. Histamine-immunoreactive endocrine cells were found almost exclusively in the acid-producing part of the stomach; only rarely were such cells observed in the pyloric gland area. They were fairly numerous in the gastric mucosa of the two subclasses of fish as well as in the amphibia and reptile species studied. Here, the majority of the histamine-immunoreactive endocrine cells seemed to have contact with the gastric lumen (open type cells) and were located in the surface epithelium (certain fish only) or together with mucous neck cells at the bottom of the pits. In the chicken, histamine-immunoreactive endocrine cells were numerous and located peripherally in the deep compound glands. They were without contact with the lumen (closed type) and had long basal extensions ("paracrine" appearance), running close to the base of the oxyntic-peptic cells. In mammals, the number of histamine-immunoreactive endocrine cells in the stomach varied greatly. They were particularly numerous in the rat and notably few in the dog, monkey and man. In all mammals, the histamine-immunoreactive endocrine cells were of the closed type and located basally in the oxyntic glands. They often had a "paracrine" appearance with long basal processes. Histamine-storing mast cells, finally, were few in both subclasses of fish as well as in the amphibian species and in the lizard. They were fairly numerous in chicken proventriculus (beneath the surface epithelium), few in the oxyntic mucosa of mouse, rat and hamster, moderate in number in hedgehog, guinea-pig, rabbit, pig and monkey, and numerous in cat, dog and man.(ABSTRACT TRUNCATED AT 400 WORDS)

APUD Cells↗
PubMed1976

Ethanol and isopropanol effects on schedule-controlled responding.

The effects of ethanol and isopropanol were studied on responding by pigeons under multiple fixed-ratio (FR), fixed-interval (FI) schedules of food presentation and under a fixed-interval (FI) schedule of food presentation where responding was decreased by punishment. The ethanol was rapidly absorbed into blood and decreased responding within 15 min after intubation to the opening of the proventriculus. Dose-effect determinations of the effects of ethanol showed that ethanol decreased responding in both the FR and FI components of the multiple schedules at similar doses, but there were increases in responding under an FR 100 schedule at lower doses. Isopropanol tended to decrease FR responding at doses that either increased FI responding or did not affect FI responding. Both ethanol and isopropanol (1 g/kg) produced effects on the local rates of responding within the FI which were rate-dependent in that they increased low rates while not affecting or actually decreasing the high rates of responding. Both ethanol and isopropanol increased punished responding if it was not severely suppressed by the punishment procedures.

1-Propanol↗
PubMed1995

Muscular architecture and VIP-like immunoreactive nerves in the gastroduodenal junction of the chicken.

The muscular architecture and the distribution of nerves with vasoactive intestinal polypeptide-like (VIP-like) immunoreactivity were examined in the gastroduodenal junction of the chicken. Both the outer and the inner muscle layers (tunica muscularis) and the muscularis mucosae were examined. The inner muscle layer was divided into two parts, namely its inner and outer parts. The outer muscle layer was not present in the pyloric part but was found in the proventriculus and the duodenum. The muscle strands of the outer muscle layer ran longitudinally but were absent at the isthmus and the pylorus. Most of the outer part of the inner muscle layer was located in or close to the pyloric part. The muscle strands in the outer part connected the pyloric part with other regions, namely the craniodorsal muscle, the tendinous centres and the isthmus. The inner part of the inner muscle layer was widely distributed in the stomach. In the pyloric part, the muscle strands of this layer followed a circular or spiral course. The muscularis mucosae was not present in the caudodorsal and cranioventral muscles of the gizzard. In addition, VIP-like immunoreactive nerve fibres were predominantly distributed in the inner part of inner muscle layer of the pyloric part. These results suggest that the ventriculopyloric orifice and the pyloric part itself constrict with a sphincter-like action.

Animals↗
PubMed2001

Chlorpyrifos-induced hsp70 expression and effect on reproductive performance in transgenic Drosophila melanogaster (hsp70-lacZ) Bg9.

Expression of hsp70 in the third-instar larval tissues of transgenic Drosophila melanogaster (hsp70-lacZ) following dietary exposure to organophosphate insecticide chlorpyrifos for various time intervals was investigated. Effect of the chemical on different developmental stages of the fly was also evaluated by looking at survivorship, hatchability, emergence, fecundity, fertility, and reproductive performance. Our results showed that the toxicant evokes profound cytotoxic effect as evidenced by dark blue staining in salivary gland, proventriculus, brain ganglia, and midgut in the lowest concentration of the chemical following 24 and 48 h of exposure. On the other hand, a significant increase (61%) in hsp70 expression in the above larval tissues was observed in the next higher concentration of the toxicant after 6 h exposure when compared with that of the lowest dose. A further increase in exposure time caused 100% larval mortality. Similarly, larvae exposed to higher concentrations of the toxicant, exhibited 100% mortality within 1 h of treatment. The insecticide caused a delay in emergence and a severe reduction in survivorship of the flies in a dose-dependent manner with 100% mortality within a day of exposure in the highest dosed group. A delay in emergence by 3 days was evident even in the lowest concentration of the chemical. A drastic effect of the chemical on hatchability was found in the highest dosed group with 100% embryonic mortality at post-16 stage of the embryo. Reproductive performance was significantly affected even in the lowest dosed group. The present study suggests that certain larval tissues of Drosophila, a nontarget organism, are vulnerable to chlorpyrifos as evidenced by hsp70 expression. Further, the adverse effect of the toxicant is reflected on various stages of development of the fly including reproductive performance.

Animals↗
PubMed2005

In vitro excystment of the metacercariae of Ribeiroia ondatrae.

In vitro excystation studies were done on the metacercarial cysts of Ribeiroia ondatrae obtained from naturally infected leopard frogs, Rana pipiens. Studies were done in Petri dish cultures in various media maintained at 41 degrees C. The media consisted of 0.5% trypsin-0.5% bile salts in Earle's balanced salt solution at pH 7.8 (TB), 1% acid pepsin (AP) in 0.85% saline adjusted to pH 2 with HCl; acid saline (AS) adjusted to pH 2 with HCl, and saline (S). The TB medium, used successfully to excyst the metacercariae of numerous species of digeneans, did not induce excystation of Ribeiroia ondatrae. Some excystation occurred in AS but not in S. Excystation was 100% in AP with most organisms excysting within 30 s after placement in the medium at 41 degrees C. Excystation in vitro in AP at 41 degrees C is compatible with the fact that this organism establishes itself in the acid-pepsin environment of the proventriculus of avian hosts.

Animals↗
PubMed1997

Laminin immunoreactivity in enteric ganglia of the chick embryo.

The localization and time of appearance of laminin in the duodenum of the chick embryo were studied with an anti-laminin polyclonal antibody and immunofluorescence. Laminin immunoreactivity was observed in the basement membranes of the mesothelium, mucosal epithelium, muscle cells and in the adventitia and basal surface of the endothelium in blood vessels. In addition, laminin immunostaining was detected over the contour of myenteric ganglia from embryonic day 7 and inside these ganglia from embryonic day 13. In colocalization experiments, laminin immunoreactivity occurred outside tubulin immunoreactive neuronal cell bodies, thus indicating that it resides in glial cells or in extracellular spaces. In addition connecting strands of the myenteric plexus and intramuscular nerves expressed laminin immunoreactivity. Similar observations were made in the proventriculus, gizzard, ileum and rectum of chick embryos, and in the duodenum and rectum of quail embryos. In the ganglion of Remak, laminin immunofluorescence was detected in the collagenous sheath that surrounds the ganglion and inside the ganglion, where it outlines neuronal cell bodies. Laminin immunoreactivity within the myenteric ganglia during the 3rd week in ovo, appears to be characteristic of the avian species examined, since it was not observed in the rat and mouse intestine at equivalent developmental stages. Immunocytochemical experiments at the electron-microscope level confirmed that structures with laminin or laminin-like immunoreactivity occur both around and inside myenteric ganglia. It is suggested that laminin, or an immunologically similar molecule, may play a role in the development and maturation of avian enteric ganglia.

Animals↗